RNA Extraction

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Questions related to RNA Extraction

How biological material (Animal carcasses, excrements) collected in field can be best stored for a later DNA and RNA extraction?

How biological materials (Animal carcasses, excrements) collected in field can be best stored for a later DNA and RNA extraction? what is the protocol to follwo and what are the storing...

26 April 2020 5,949 11 View

How do we calculate cumulative mortality % and Relative percentage survival (RPS) for fish challenge studies?

I wanted to know how to calculate RPS and CM% with detailed explanation of the same. I also wanted to know how do we express this data?

20 April 2020 2,220 0 View

Has anyone had experience in simultaneous extraction of mRNA, miRNA, and LncRNAs?

Hello everyone, I want to measure the expression change of some mRNA, miRNA, and LncRNA at once under a specific condition. Since I am going to do relative quantification, is there any RNA...

23 March 2020 422 7 View

Troubleshoot help with qPCR and protein extraction?

Hello I am working on a project where I need to extract RNA from my samples. For some reason, Nanodrop gives me values below [10 ng/uL] for almost all my samples, even though I saw a pellet in...

04 March 2020 995 4 View

Is there any alternative to a fume hood when working with DTT?

I am going to work with dithiothreitol (DTT) whilst isolating DNA and RNA, so I was looking for a safe alternative to a fume hood, such as maybe a specific type of mask or anything different that...

18 February 2020 3,946 3 View

How to create "trait data" file for WGCNA?

As a biotechnology student in Shahid Beheshty University, Tehran Iran, I am very interested in systems biology area. Actually I need to know how should I create trait matrix file to reach Heatmap...

16 February 2020 1,892 3 View

TRIzol RNA extraction: No pellet after isopropanol precipitation even with glycogen?

So I'm using TRIzol for RNA extraction, but suddenly I'm getting no pellet during the isopropanol step *even though I added glycogen.* A few weeks ago I: 1. Took a large quantity of bacteria 2....

06 January 2020 8,556 5 View

Why is there no clear interphase during Trizol RNA extraction of fibroblast cells?

It looks almost as if the interphase is floating up into the aqueous phase during my extraction procedure. It doesn't form a distinct layer; it is very irregularly shaped and juts out into the...

02 January 2020 4,819 4 View

Reliability of Gel or Nanodrop ratios for RNA integrity and quality?

I have read somewhere that these should be only taken as a frame of reference? I am assuming that my samples with a 260/280 ratio of ~1.5 are too low, but when I run it on a 1% agarose gel, the...

26 December 2019 3,736 3 View

Who can help me in the dentification of such strange structures in common sole stomach contents?

Hi everyone We've found such strange structures in a common sole (Solea solea) stomach from the Northern Adriatic. They look like "calcified scales", with growth rings-like features Someone can...

13 December 2019 2,941 2 View

Why i am not getting the 2 layers/phases after adding 100% ethanol?

In isolation of DNA from trizol method, after adding the absolute ethanol in the phase left behind (RNA Extraction step) i am not getting the 2 layers to proceed further for DNA extraction and...

12 November 2019 1,567 2 View

Compatible of QIAseq Stranded Total RNA Lib Kit and Ribominus Transcriptome Isolation Kit?

Dear all, I'm going to perform RNA-Seq with a QIAseq Stranded Total RNA Lib Kit and depletion with Ribominus Transcriptome Isolation Kit/ Ribominus concentration Kit. Does anyone have...

29 October 2019 8,312 1 View

Low bacterial RNA yeld with RNeasy MiniKit?

Dear all, I am trying to do an RNA extraction of different bacterial species with RNAeasy mini kit, protokol #4 (lysozyme lysis + proteinase K digestion). Unfortunately, I have a fairly low...

28 October 2019 5,729 0 View

How can I improve the RNA extraction from cells?

Hello, I have problems with RNA extraction from cells. I work with primary neurons and astrocytes cultures, with 1*10^6 cells for each well (more or less, because some cells die). Actually, I use...

02 October 2019 1,701 5 View

How much cq value and error can be acceptable for reference and targeted gene in qPCR?

Dear Researchers, I did the qPCR, 20ul, (10 ul TB gree, primer 0.4, template 2ul, Water 7.2), steps, by using TAKARA, I got the following results, as you can see that with 1X, 3X, and 5X...

29 September 2019 4,437 9 View

What should do, if RNA quality is very low?

Dear Researcher I am working on passion fruit peel at ripening stages, and want to do qPCR but facing problem in getting good quality RNA from fruit peel, I have tried to extract the RNA...

28 September 2019 1,971 3 View

Which protocol for RNA Extraction from bacterial culture growing on aromatic compounds (problems with dark precipitations/polyphenolic compounds)?

During the course of the cultivation of a Pseudomonas sp. strain on a toluene derivative we observe a strong darkening of the media, supposedly because of polyphenolic (polycatecholic) compounds...

27 September 2019 9,923 6 View

RNA extraction from FAA fixed, methanol-preserved plant tissue. Is it possible?

Dear colleagues, I have a bucket of carrot's inflorescence buds that were fixed with FAA, dehydrated and stored in methanol at -20°C for ca. 3 months. I did ISH with some of them that worked fine...

23 September 2019 7,897 2 View

Can i use Tween-20 instead NP-40/Triton X-100 for separation of cytoplasmic and nuclear RNA?

Hi there. I want to extract separately cytoplasmic and nuclear RNA fraction from leukemic cell line by Trizol. Most of protocols that I saw suggests using a buffer with NP-40 or Triton X-100 for...

07 September 2019 2,626 3 View

What is the ideal timeline for differentiating Mono-Mac 1 cells with LPS?

We are currently planning experiments with the monocytic cell line Mono-Mac 1, which we will be using for HIV infections and analysis of different signaling pathways. As we are using them as a...

06 September 2019 3,479 2 View

What are the calculations to make a constant RNA concentration for different samples to do cDNA SYNTHESIS?

I have 2 concentrations from different samples, Nanodrop for each are 715.5 ng/ul and 467.5 ng/ul and approximately 2 ul was used for each sample. I am very confused on how to get and equal...

23 August 2019 3,665 3 View

What is the most credible way to quantify temporal functional-connectivity among EEG channels?

I am searching for a time-dependent measure of temporal functional connectivities between different pair of channels so the result is also a time series not a scaler. and also possibly a good way...

07 August 2019 7,423 5 View

How to extract RNA from cell cultured on hybrid scaffold composed of PCL and collagen?

Hi! We are doing studies about tissue engineering using hybrid scaffold composed of PCL and collagen. However get stuck in RNA extraction for gene expression analysis. Some study suggest using the...

19 July 2019 1,012 3 View

Why I lose my lettuce hairy roots RNA after extraction with licl method?

Why I lose my lettuce hairy roots RNA about 5 or 6 hours after extraction with licl method and on electrophoresis only it's DNA band will remain with no RNA? I do the extraction and I get the RNA...

10 July 2019 9,063 2 View

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